(1) Field of the Invention
The present invention relates to methods of purification, immobilization and amplification of nucleic acids. Specifically, nucleic acid amplification using a solid support.
(2) Description of Related Art
There are a variety of methods for the purification and amplification of nucleic acids known to those skilled in the art. However, these methods can be slow, tedious, expensive and difficult to automate and manufacture. They also lack the performance in regards to sensitivity, specificity, precision, accuracy and other features that would otherwise allow them to be used successfully in a number of diverse applications. These methods are often complicated, requiring isolation of nucleic acid targets from samples, preparing the targets for amplification, performing amplification and detecting the amplification product. In some applications, this may also include preparing templates for sequencing including next generation sequencing and sequencing the target nucleic acids. In many of these methods one or more purification steps may also be required. All of these steps increase performance time, cost, complexity and labor. Providing methods with versatility as to specificity, sensitivity, selectivity, precision and accuracy is important. As are methods that are rapid, efficient and easy to use with minimal assay steps and reagents for readily obtaining a desired isolated and amplified product from a raw sample. These products may be amplified nucleic acids or set of amplified nucleic acids (i.e., multiplexing, including high level multiplexing) that may be utilized as templates, free in solution or on solid supports, for immediate sequencing and next generation sequencing. In addition, methods for directional amplification that preserve strand orientation provide proper alignment of sequences during mapping and more accurate quantitative determinations (e.g. of gene expression levels). In addition, these methods can be utilized with a wide variety of nucleic acid amplification methods that are known in the art. In addition these methods can be utilized with tag sequences. The present invention provides methods for purifying, immobilizing and amplifying specific nucleic acids from a sample that overcome many of the current limitations in the art.